1、BRITISH STANDARDBS ISO 23893-1:2007Water quality Biochemical and physiological measurements on fish Part 1: Sampling of fish, handling and preservation of samplesICS 13.060.70g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3
2、g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58BS ISO 23893-1:2007This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 December 2007 BSI 2007ISBN 978 0 580 53705 9National forewordThis British Standard is the UK impl
3、ementation of ISO 23893-1:2007.The UK participation in its preparation was entrusted by Technical Committee A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users
4、 are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations. Amendments issued since publicationAmd. No. Date CommentsEH/3, Water quality, to Subcommittee EH/3/5, Biological methods.Reference numberISO 23893-1:2007(E)INTERNATIONAL STA
5、NDARD ISO23893-1First edition2007-11-15Water quality Biochemical and physiological measurements on fish Part 1: Sampling of fish, handling and preservation of samples Qualit de leau Mesurages biochimiques et physiologiques sur poisson Partie 1: chantillonnage des poissons, manipulation et conservati
6、on des chantillons BS ISO 23893-1:2007ii iiiContents Page Foreword iv Introduction v 1 Scope . 1 2 Principle. 1 3 Equipment . 2 4 Fish sampling 3 4.1 Statistical aspects. 3 4.2 Frequency and season for sampling 3 4.3 Selection of sampling sites . 3 4.4 Sampling procedures . 4 4.5 Handling of samples
7、 and analytical procedures . 6 4.6 Background information 7 5 Quality assurance. 7 5.1 General. 7 5.2 Fish sampling 7 5.3 Tissue sampling 7 5.4 Biochemical/chemical analysis . 7 5.5 Evaluation 7 6 Report 8 6.1 General. 8 6.2 Data logging, data hosting. 8 6.3 Evaluation 8 Annex A (informative) Summar
8、y of variables used as biomarkers in fish . 9 Annex B (informative) Guide to interpretation of biomarker responses with references. 12 Annex C (informative) Suggested report for fish sampling . 15 Annex D (informative) Suggested report for tissue sampling . 16 Bibliography . 18 BS ISO 23893-1:2007iv
9、 Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a
10、technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matter
11、s of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulat
12、ed to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible
13、 for identifying any or all such patent rights. ISO 23893-1 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods. ISO 23893 consists of the following parts, under the general title Water quality Biochemical and physiological measurements on fish: Part
14、1: Sampling of fish, handling and preservation of samples Part 2: Determination of ethoxyresorufin-O-deethylase (EROD) Technical Specification BS ISO 23893-1:2007vIntroduction Determination of biomarker responses can be used to detect toxicity of known as well as unknown pollutants, when they occur
15、singly or in combination. Therefore, measurement of biomarkers is a cost-effective way to assess ecosystem health. In combination with determinations of occurring and suspected pollutants, determinations of biomarkers can facilitate the interpretation of cause-effect relationships in the environment
16、, as well as in laboratory toxicity tests. Information on commonly used biomarkers and the interpretation of biomarker responses is given in Annexes A and B, respectively. Biomarkers like ethoxyresorufin-O-deethylase (EROD), metallothionein and vitellogenin are used to detect and quantify sublethal
17、effects of pollutants, especially in fish. However, many of the biochemical and physiological variables that are used as biomarkers are sensitive not only to disturbances by the pollutants of concern, but also by the normal biochemical and physiological adjustments made by the fish in response to se
18、asonal variation, its normal development and sexual maturation. Some variables can also be affected by general stress to disturbances caused by the handling during fish and fish tissue sampling. Therefore, standardisation of procedures used for sampling and handling of samples prior to determination
19、 of the biochemical and physiological variables is important. Sublethal responses at the individual level usually occur before effects are seen at the population and community level. In the aquatic environment, fish are suitable for detection of physiological effects of pollutants, because they are
20、exposed both through the water and through their food organisms. Also, the physiology and biochemistry of fishes is rather similar to that of humans and other vertebrates, making comparisons with studies on mammals easier than for those with crustaceans and other invertebrates. This part of ISO 2389
21、3 serves as guidance for sampling and a platform for determination of biomarkers in fish, making it possible to use the measurements to: describe the state of the environment regarding effects of anthropogenic compounds on the health of fish; perform time-trend surveillance (monitoring); provide ref
22、erence data and material for assessment of effects from point sources; evaluate and assess environmental threats; provide background information for environmental measures; follow up and assess effects of environmental corrective measures; integrate the biomarker responses with other measurements (e
23、.g. fish abundance, recruitment and pollutant residues) in order to facilitate the interpretation of environmental status or impact. BS ISO 23893-1:2007blank1Water quality Biochemical and physiological measurements on fish Part 1: Sampling of fish, handling and preservation of samples 1 Scope This p
24、art of ISO 23893 provides guidance on how to sample fish for determination of biochemical and physiological characteristics, such as the composition and enzyme activities of blood, liver, muscle and other tissues in order to asses the health of fish in the field as well as in the laboratory. The bio
25、chemical and physiological variables used for this purpose are often called biomarkers. This part of ISO 23893 includes recommendations and methods for: obtaining a site-specific sample of a representative number of fish; sampling fish tissues in the field and in the laboratory; and handling and pre
26、servation of samples prior to analysis of biochemical and physiological variables. 2 Principle Fish of a suitable species, age (size), and sex are sampled at selected sites at a suitable time of the year in order to reduce variability due to biological, geographical, and seasonal influences. Standar
27、dised sampling and measurement procedures, and qualified staff are used for collection of samples, transport, storage, and analysis. By these means, the results from time series of comparable data can be used to detect changes in the environment that are caused by anthropogenic compounds. Necessary
28、permits for fish and fish tissue sampling shall be obtained to comply with national legislation. This may include permits from the (land) owner of the fishing rights, regional environmental and fishery authorities, and ethical (animal rights) authorities. The health of fish can be assessed by determ
29、ination of biochemical, physiological, histological, and pathological methods. The subcellular and cellular variables are often called biomarkers. The primary toxic effect of pollutants, which occurs at the subcellular level, results in a biochemical or physiological change. This reaction is usually
30、 fast, and it can progress further and cause disturbances at higher levels of biological organisation within the organism, resulting in changes at the cellular and tissue (organ) level (histological changes). These can lead to disturbances of reproduction and growth, and can eventually cause death o
31、f the organism. Monitoring of fish health can, therefore, serve as an early warning system for anthropogenic disturbance. Through a combination with other measurements (integrated monitoring), it may be possible to correlate biomarker responses with for instance pollutant residues, distance from poi
32、nt sources, and ecological variables such as reproductive recruitment, which are known to be sensitive to pollutants. In principle, this method can be applied to all species of fish from all types of environments (fresh, salt, brackish, cold, and warm water) and in shallow as well as reasonably deep
33、 water habitats. However, it is usually advantageous to restrict these methods to certain species of fish, which can be used as indicator species for fish health. These species shall be stationary, readily available (catchable in most locations) and reasonably resistant to handling stress. Their bio
34、logy and physiology should be well known in order to make the interpretation of data easier. Examples of such species are the perch (Perca fluviatilis) and the eelpout (viviparous blenny, Zoarces viviparus), which are used for monitoring along the Swedish coast. BS ISO 23893-1:20072 Preferably the f
35、ish species used in the field should be suitable for keeping in the laboratory for toxicological studies to investigate and confirm cause-effect relationships detected or suspected to take place in the field. Procedures for organ and tissue sampling are essentially identical in both field and labora
36、tory studies. Procedures for collection of fish for field studies and for collection of organs are, therefore, described in separate sections. 3 Equipment 3.1 Fish sampling equipment 3.1.1 Fishing boat, suitable for the area. 3.1.2 Clothing for outdoor work. 3.1.3 Lifejacket, of suitable size and bu
37、oyancy for each crew member. 3.1.4 Gill nets, made from textile or nylon fibres and of specified and suitable size for catching the desired species and size and their gentle release into the fish chest used for storage. 3.1.5 Other equipment for fish capture, e.g. electroshocker and fyke nets, shall
38、 be described in enough detail to allow interpretation and repeated sampling. 3.1.6 Global positioning system (GPS) instrument, for exact location of sampling sites. 3.1.7 Nautical map, for marking of sampling sites. 3.1.8 Knife and pair of scissors, for gentle removal of fish from gill nets. 3.1.9
39、Fish chest, made from wood or other inert material for storage of fish before tissue sampling. 3.1.10 Instruments for measurement of physical and chemical characteristics of water, e.g. thermometer, pH meter, conductivity meter. 3.1.11 Equipment for determination of water depth, an echo-sounder or a
40、 calibrated line can be used to determine the depth. 3.2 Tissue sampling equipment 3.2.1 Jetty, with easy access to fish chest and within 100 m of the field laboratory. 3.2.2 Landing net, suitable for the fish species and size. 3.2.3 Field laboratory, boathouse, garage or mobile laboratory supplied
41、with electricity. 3.2.4 Stick (baton), for stunning the fish prior to sampling of blood. 3.2.5 Anaesthetic, to anaesthetise the fish (details on usage are given in 4.4.3). 3.2.6 Dissection equipment: forceps, scissors, scalpel, syringes, needles. 3.2.7 Ruler, for determination of body length. 3.2.8
42、Balance, for determination of body mass and tissue (liver, gonad, spleen) mass. 3.2.9 Centrifuge and tubes, for blood plasma. 3.2.10 Microscope slides, for preparation of blood smears. BS ISO 23893-1:200733.2.11 Sample containers, of suitable sizes for tissue samples (e.g. of plastic with snap locks
43、). 3.2.12 Marking pen, waterproof and freezeproof. 3.2.13 Vacuum flask with liquid nitrogen, for rapid freezing and temporary storage of samples. 3.2.14 Container with solid carbon dioxide, for transport of deep-frozen tissue samples between field laboratory and analytical laboratory. 3.3 Biomarker
44、determination equipment for the field laboratory 3.3.1 Haematocrit tubes and centrifuge, if required. 3.3.2 Blood glucose meter, if required. 3.3.3 Haemoglobin meter, if required. 4 Fish sampling 4.1 Statistical aspects Feral fish, like other wild animals, are affected by a number of natural factors
45、 besides those caused by anthropogenic load. Important natural factors for fish are climate, hydrology, oxygen and salinity (abiotic factors), as well as age, size, sex, maturation, nutritional status, parasites and diseases (biotic factors). All these factors can contribute to the overall variabili
46、ty of the measured response variables. In order to detect temporal changes in trend monitoring and geographical variation in mapping of potential disturbance, all the abiotic and biotic factors mentioned above shall be reduced in importance as much as possible. 4.2 Frequency and season for sampling
47、Fish should be sampled once a year during the autumn period in order to avoid the effects of rapid changes in physiological conditions due to the reproduction season. During the autumn, most species of fish are not reproducing, and the conditions to get enough fish by stationary gear like gill nets
48、(3.1.4) and fyke nets (3.1.5) are still good because the fish are still active. More frequent sampling at other times of the year does generally not provide any new information in trend monitoring. In Sweden, perch for fish-health monitoring is sampled by gill nets in September, and eelpout by fyke
49、nets in November. The most suitable period differs between countries and regions due to differences in climate. Often only sexually mature fish of one sex (e.g. females for perch and eelpout, and males for chub and zebrafish) within a certain size interval are used for each species in order to minimise the influence of sex and size. 4.3 Selection of sampling sites In fish-health monitoring, it is of utmost importance to have as much detailed information as possible about the anthropogenic load on sites to be used as reference locations. These sites should be monitor
